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[Purification of hsp70 and its immunoprotective effect against mouse hepatoma]
1Liver Cancer Institute, Zhongshan Hospital, Shanghai Medical University, Shanghai 200032, China.
Objective:
To purify hsp70 protein and observe its anti-tumor effect on mouse H22 hepatoma.
Methods:
Cytosolic hsp70 protein of heat treated H22 cells was purified successively by chromatographic procedures, including DEAE 52-cellulose, ConA-sepharose 4B and ADP-argarose chromatography. The molecular weight and the identity of the purified hsp70 protein were confirmed by SDS-PAGE and Western blot. Tumorigenicity of H22 hepatoma was examined in purified hsp70-treated C3H mice.
Results:
A sharp stained protein band with a molecular weight of about 70 kD was obtained and shown to be hsp70 as confirmed by Western blot. In C3H mice challenged with H22 hepatoma, tumor developed in 6 of 6 mice while in hsp70 pretreated mice, none of the 6 H22 challenged mice developed tumor. Nevertheless, C57BL/6 mice pretreated with hsp70 or RPMI 1640 all developed tumor upon challenge with EL-4 lymphoma.
Conclusion:
The results suggest that the immunoprotective effect of hsp70 protein is due not to hsp70 per se, but rather to the specific antigenic peptides it carries.